Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China.
Tianjin Hospital, Tianjin, China.
Mol Vis. 2022 Dec 21;28:441-450. eCollection 2022.
To investigate the therapeutic effect of lipoxin A4 (LXA4) on -stimulated human corneal epithelial cells (HCECs).
The cell counting kit-8 (CCK-8) was performed in HCECs to evaluate the toxicity of LXA4. A cell scratch test was used to assess the impact of LXA4 on the migration of HCECs. Enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), and western blot were applied to examine the expression of inflammatory mediators in -stimulated HCECs. The nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear translocation and expression in HCECs were detected by immunofluorescence staining.
LXA4 at 0-10 nmol·L (nM) had no significant cytotoxic effect on HCECs. LXA4 at a concentration of 1 nM and 10 nM significantly promoted the migration rate of HCECs. The mRNA and protein levels of pro-inflammatory mediators, including IL-1β, TNF-α, and IL-6, were remarkably lower in the LXA4-treated group. LXA4 promoted the expression of Nrf2 and heme oxygenase 1 (HO-1) in -stimulated HCECs compared with the PBS control group. Pretreatment with brusatol (BT, Nrf2 inhibitor) or Zine Protoporphyrin (Znpp, HO-1 inhibitor) receded the anti-inflammatory ability of LXA4.
LXA4 plays a protective role in -stimulated HCECs by inhibiting the expression of pro-inflammatory mediators through the Nrf2/HO-1 signaling pathway.
研究脂氧素 A4（LXA4）对脂多糖（LPS）刺激的人角膜上皮细胞（HCECs）的治疗作用。
采用细胞计数试剂盒-8（CCK-8）检测 LXA4 对 HCECs 的毒性。细胞划痕实验评估 LXA4 对 HCECs 迁移的影响。酶联免疫吸附试验（ELISA）、实时定量聚合酶链反应（qRT-PCR）和蛋白质印迹法检测 LPS 刺激的 HCECs 中炎症介质的表达。免疫荧光染色检测核因子红细胞 2 相关因子 2（Nrf2）在 HCECs 中的核转位和表达。
0-10 nmol·L（nM）的 LXA4 对 HCECs 无明显细胞毒性作用。1 nM 和 10 nM 的 LXA4 显著促进 HCECs 的迁移率。LXA4 处理组促炎介质（包括 IL-1β、TNF-α 和 IL-6）的 mRNA 和蛋白水平显著降低。与 PBS 对照组相比，LXA4 促进了 LPS 刺激的 HCECs 中 Nrf2 和血红素加氧酶 1（HO-1）的表达。用 Brusatol（BT，Nrf2 抑制剂）或锌原卟啉（Znpp，HO-1 抑制剂）预处理可降低 LXA4 的抗炎作用。
LXA4 通过 Nrf2/HO-1 信号通路抑制促炎介质的表达，在 LPS 刺激的 HCECs 中发挥保护作用。
